Predominance of Leishmania (Leishmania) amazonensis DNA in Lutzomyia longipalpis sand flies (Diptera: Psychodidae) from an endemic area for leishmaniasis in Northeastern Brazil

ABSTRACT Leishmaniasis is a serious public health concern in the Northeastern region of Brazil, where the sand fly fauna is well studied, although few species have been identified as competent vectors. The detection of Leishmania spp. parasites in wild-caught sand flies could help sanitary authorities draw strategies to avoid the transmission of the parasites and, therefore, the incidence of leishmaniases. We detected Leishmania DNA in wild-caught sand flies and correlated that data with aspects of sand fly ecology in the Caxias municipality, Maranhao State, Brazil. The sand flies were sampled in the peridomicile (open areas in the vicinity of human residences) and intradomicile (inside the residences) from July/2019 to March/2020. Leishmania DNA was detected in females, targeting a fragment of the Internal Transcribed Spacer (ITS1) from ribosomal DNA. Among the fourteen species of sand flies identified, five (Lutzomyia longipalpis, Nyssomyia whitmani, Evandromyia evandroi, Micropygomyia trinidadensis, and Micropygomyia quinquefer) harbored DNA of Leishmania (Leishmania) amazonensis. The most abundant species in rural (Ny. whitmani: 35.2% and Ev. evandroi: 32.4%) and urban areas (Lu. longipalpis: 89.8%) are the permissive vectors of L. (L.) amazonensis, especially Ny. whitmani, a known vector of causative agents of cutaneous leishmaniasis. Although Lu. longipalpis is the vector of L. (L.) infantum, which was not detected in this study, its permissiveness for the transmission of L. (L.) amazonensis has been reported. We suspect that visceral leishmaniasis and cutaneous leishmaniasis are caused by L. (L.) amazonensis, and the transmission may be occurring through Lu. longipalpis, at least in the urban area.

Integration of phlebotomine ecological niche modelling, and mapping of cutaneous leishmaniasis surveillance data, to identify areas at risk of under-estimation.

INTRODUCTION: Passive surveillance systems are thought to under-estimate the true incidence of American cutaneous leishmaniasis (ACL) by two- to five-fold. Ecological niche models based on remotely sensed data can identify environmental factors which favor phlebotomine vectors. Here we report an integrated approach to identifying areas at risk of cutaneous leishmaniasis by applying spatial analysis methods to niche model results, and local surveillance data, in two locations in Colombia with differing vector ecology. The objective was to identify townships in which later phases of the project could implement community-based surveillance to obtain direct estimates of under-reporting. MATERIALS AND METHODS: The study was carried out in one municipality in each of two departments of the Andean region of Colombia: Pueblo Rico in Risaralda, and Rovira in Tolima. Niche mapping by maximum entropy, based on published and unpublished existing locations of Pintomyia (Pifanomyia) longiflocosa and Psychodopygus panamensis, and using variables on land cover, climate and elevation. Field catches were done in each municipality to test predictions of high relative probability of presence. The niche model results were included as a predictor in a conditional autoregressive spatial model, in which the outcome variable was the number of cases by township, as detected by passive surveillance. RESULTS: Having rarefied 173 geolocated records, 46 of Pi. longiflocosa and 57 of Ps. panamensis were used for the niche modelling. At the national level, both species had high relative probability of presence on parts of the slopes of the three Andean cordilleras. Pi. longiflocosa also has a high relative probability of presence in the higher parts of the Magdalena valley, as does Ps. panamensis in some areas close to the Caribbean coast. At the local level, field catches confirmed that Pi. longiflocosa was the most abundant species in Rovira, and likewise Ps. panamensis in Pueblo Rico. The spatial regression showed that the incidence of ACL, according to surveillance, was positively, but not statistically significantly, associated with the relative probability of presence from the risk model. CONCLUSIONS: These niche maps bring together published and unpublished results on phlebotomine species which are important vectors in Colombia. Maps of the fitted values of incidence were used to guide the selection of townships in which further phases of the study will attempt to quantify the extent of under-estimation of ACL incidence.

Urban Horses As Environmental Bioindicators for Leishmaniasis.

The presence of DNA and anti-Leishmania spp. antibodies in the serum of 112 healthy horses was investigated by evaluating the physical examination, from a rural society located in the north central region of Paraná. The antigens of Leishmania amazonensis, Leishmania braziliensis, and Leishmania infantum were used to perform the indirect enzyme-linked immunosorbent assay, where it was possible to detect the reaction in 27.67% of the samples. These were also subjected to the real-time quantitative PCR, which confirmed the presence of Leishmania spp. DNA in 67.34% of the tested samples. The results show that the tested animals were previously exposed to the protozoan. Thus, these animals can be considered environmental bioindicators of the presence of Leishmania spp. at the study site. The material used in this study (serum), although not ideal, proved to be effective and less invasive. Taking into account the importance of the disease, the absence of more in-depth information on the species, the high zootechnical value of these animals, and their strictly close contact with the urban area and the human species, it is essential that further studies are carried out to elucidate the epidemiological profile of them in the face of the disease, as well as the possibility of them acting not only as hosts but also as reservoirs.

Potential vectors of Leishmania spp. in an Atlantic Forest conservation unit in northeastern Brazil under anthropic pressure.

BACKGROUND: Phlebotomines are a group of insects which include vectors of the Leishmania parasites that cause visceral leishmaniasis (VL) and cutaneous leishmaniasis (CL), diseases primarily affecting populations of low socioeconomic status. VL in Brazil is caused by Leishmania infantum, with transmission mainly attributed to Lutzomyia longipalpis, a species complex of sand fly, and is concentrated mainly in the northeastern part of the country. CL is distributed worldwide and occurs in five regions of Brazil, at a higher incidence in the north and northeast regions, with etiological agents, vectors, reservoirs and epidemiological patterns that differ from VL. The aim of this study was to determine the composition, distribution and ecological relationships of phlebotomine species in an Atlantic Forest conservation unit and nearby residential area in northeastern Brazil. METHODS: Centers for Disease Control and Shannon traps were used for collections, the former at six points inside the forest and in the peridomestic environment of surrounding residences, three times per month for 36 months, and the latter in a forest area, once a month for 3 months. The phlebotomines identified were compared with climate data using simple linear correlation, Pearson's correlation coefficient and cross-correlation. The estimate of ecological parameters was calculated according to the Shannon-Wiener diversity index, standardized index of species abundance and the dominance index. RESULTS: A total of 75,499 phlebotomines belonging to 11 species were captured in the CDC traps, the most abundant being Evandromyia walkeri, Psychodopygus wellcomei and Lu. longipalpis. Evandromyia walkeri abundance was most influenced by temperature at collection time and during the months preceding collection and rainfall during the months preceding collection. Psychodopygus wellcomei abundance was most affected by rainfall and relative humidity during the collection month and the month immediately preceding collection time. Lutzomyia longipalpis abundance showed a correlation with temperature and the rainfall during the months preceding collection time. The Shannon trap contained a total of 3914 phlebotomines from these different species. Psychodopygus wellcomei, accounting for 91.93% of the total, was anthropophilic and active mainly at night. CONCLUSIONS: Most of the species collected in the traps were seasonal and exhibited changes in their composition and population dynamics associated with local adaptions. The presence of vectors Ps. wellcomei and Lu. longipalpis underscore the epidemiological importance of these phlebotomines in the conservation unit and surrounding anthropized areas. Neighboring residential areas should be permanently monitored to prevent VL or CL transmission and outbreaks.

Land use in relation to composition and abundance of phlebotomines (Diptera: Psychodidae) in five foci of domiciliary transmission of cutaneous leishmaniasis in the Andean region of Colombia.

American cutaneous leishmaniasis is a public health concern in Colombia, its incidence being sustained or focally increased principally by the emergence of domestic transmission concomitantly with the adaptation of the phlebotomine vectors to habitat transformation around households. The objective of the study is to scale up a rapid characterization methodology for evaluating the relationship of land use around the house to the composition and abundance of phlebotomines. Five sites with a history of domestic leishmaniasis transmission in the Andean area of Colombia were selected. The peri-domestic habitat was evaluated at 10 m intervals along eight radial transects, centered on each house, at 45° intervals using a web pattern. Phlebotomines were captured by placing three CDC light traps over two nights both indoors and outdoors (10 m from the house). Blood source and infection were determined by PCR. Spearman rank correlation coefficients and negative binomial regression were used to quantify associations between the phlebotomine abundance and habitat categories. The study demonstrated that the vectors were largely anthropophagic (62% of 79 were human blood) and that a single species in each site was favored by the pertaining agriculture monoculture. Specifically, Pintomyia (Pifanomyia) quasitownsendi was associated with sugar cane in Novillero and La Esmeralda; while Pi. (Pif.) longiflocosa was associated with coffee plantations in Agua Bonita and El Cucal. Honda Alta had a more diverse array of land use and forest coverage with a lower number of specimens but higher species diversity. In terms of distance from the house to an area of a given land use, the abundance of Pi. (Pif.) quasitownsendi was inversely related to the distance to sugar cane plantation (Spearman correlation coefficient, ρ = -0.56, p < 0.001 for outdoor catches, and ρ = -0.50, p < 0.001 indoors). A similar inverse relationship was observed for Pi. (Pif.) longiflocosa with regard to technified coffee (ρ = -0.51, p < 0.001 outdoors, and ρ = -0.48, p < 0.001 indoors). This rapid characterization methodology could guide public heath decision makers in identifying those houses at higher risk of domestic transmission, and also educate farmers to increase the distance between their crops and any neighboring houses.

Epidemiological aspects of Leishmaniasis in the Pantanal region of Mato Grosso / Aspectos epidemiológicos das Leishmanioses na região do Pantanal de Mato Grosso

Abstract Leishmaniasis is a disease caused by the protozoan Leishmania parasite that is disseminated by some species of sandflies and hosted by a variety of reservoirs. The objective of the present study was to evaluate the epidemiological situation of leishmaniasis in the municipalities of the Mato Grosso Pantanal. Human data were obtained from the Information System of Notifiable Diseases, and the canine and vector results from the State Department of Health of Mato Grosso. Between 2007 and 2016, 10 cases of visceral leishmaniasis and 499 cases of tegumentary leishmaniasis were identified in the Pantanal region. The surveillance studies regarding the canine reservoir demonstrated that the parasite was present in six of the seven municipalities. Vectors of visceral leishmaniasis were present in five municipalities and vectors of tegumentary leishmaniasis in six. Enhancement of services aimed at controlling this disease is fundamental to prevent an increase in the number of cases in the region.

Resumo As leishmanioses são doenças causadas por protozoários do gênero Leishmania disseminadas por algumas espécies de flebotomíneos e participação de uma variedade de reservatórios. O objetivo do presente estudo foi avaliar a situação epidemiológica das leishmanioses nos municípios abrangentes do Pantanal de Mato Grosso. Os dados humanos foram obtidos do Sistema de Informações de Agravos de Notificação e os resultados caninos e vetoriais junto à Secretaria de Estado de Saúde de Mato Grosso. A região do Pantanal registrou no período de 2007 a 2016, dez casos da forma visceral e 499 casos da forma tegumentar. As ações de vigilância para o reservatório canino demonstraram a circulação do parasito em seis dos sete municípios. A presença dos vetores de leishmaniose visceral ocorreu em cinco municípios e os da leishmaniose tegumentar em seis. O fortalecimento dos serviços voltados para controle desse agravo é fundamental para evitar o aumento no número de casos da região.

Sandflies in an endemic area for Visceral Leishmaniasis in Northeastern Brazil / Flebotomíneos em uma área endêmica para Leishmaniose Visceral no Nordeste do Brasil

Abstract Phlebotomines of the genus Lutzomyia are important vector for species of Leishmania, the etiological agent of leishmaniasis. The aim of this study was to assess the diversity of sandflies in an endemic area for Visceral Leishmaniasis (VL). Sampling was performed in four localities on the coast of the municipality of Goiana, state of Pernambuco, Northeastern Brazil. Specimens were collected monthly on three consecutive nights, from November 2017 to October 2018. CDC light traps were installed at a height of 1.5 m above the ground, in two ecotypes (i.e. tree and animal facility), both in peridomestic areas. A total of 5,205 sandflies were collected, among which the species Lutzomyia longipalpis (99.85%; 5,197/5,205) was the most common, followed by Lutzomyia evandroi (0.13%; 07/5,205) and Lutzomyia whitmani (0.02%; 01/5,205). Specimens of L. longipalpis were collected throughout the study period and most of them were found near to the animal facilities. The findings from this research indicate that vectors for Leishmania infantum are present in the study area throughout the year, along with occurrences of vectors relating to Cutaneous Leishmaniasis (CL). Preventive vector control measures need to be implemented in Goiana to reduce the risk of infection for the human and animal populations.

Resumo Flebotomíneos do gênero Lutzomyia são importantes vetores de espécies de Leishmania, agente etiológico das leishmanioses. O objetivo deste estudo foi avaliar a diversidade de flebotomíneos em área endêmica para Leishmaniose Visceral (LV). As coletas foram realizadas em quatro localidades do litoral do município de Goiana, Pernambuco, Nordeste do Brasil. Os espécimes foram coletados mensalmente em três noites consecutivas de novembro de 2017 a outubro de 2018. Foram instaladas armadilhas luminosas modelo CDC a uma altura de 1,5 m acima do solo em dois ecótopos (ex. árvore e instalações de animais), ambos em áreas peridomiciliares. Um total de 5.205 flebotomíneos foi coletado, sendo Lutzomyia longipalpis (99,85%; 5.197/5.205) a mais comum, seguida por Lutzomyia evandroi (0,13%; 07/5.205) e Lutzomyia whitmani (0,02%; 01/5.205). L. longipalpis foi coletada durante todo o período do estudo. A maioria dos espécimes foi detectada perto das instalações dos animais. Os achados deste estudo indicam a presença de vetores de Leishmania infantum na área avaliada ao longo do ano, bem como a ocorrência de vetores relacionados à Leishmaniose Cutânea (LC). Medidas preventivas de controle vetorial devem ser implementadas em Goiana para reduzir o risco de infecção à população humana e animal.

Seroepidemiological survey of canine visceral leishmaniasis in a continental island of northeast Brazil / Investigação soroepidemiológica da leishmaniose visceral canina em uma ilha continental do Nordeste do Brasil

Introduction: Visceral leishmaniasis (VL) is endemic disease in the neighboring municipalities of the continental island Ilha de Maré, Salvador, Bahia, Brazil. Nevertheless, VL has not been reported in the island itself. Objective: the present study aimed to investigate the seroprevalence and clinical signs of Canine Visceral Leishmaniasis (CVL) and to identify the sand fly population present in the village of Botelho, Ilha de Maré. Methodology: sera of local dogs were tested for anti-Leishmania IgG by immunoassays (screening with TR DPP ™ rapid test and confirmatory with indirect ELISA, Bio-Manguinhos/Fiocruz) and an entomological survey was conducted to estimate and identify the phlebotomine fauna of the region. Results: seven out of 106 samples (6.6%) were positive using rapid test. These positive samples were sent to the Central Laboratory of Bahia for confirmation by indirect ELISA. However, all samples presentednegative results. Nine specimens of Pressatia choti, subfamily Phlebotominae were identified, being this species frequently found in areas with cutaneous leishmaniasis transmission in Brazil. Conclusion: although this work did not confirm the presence of CVL in Ilha de Maré, new serological and entomological studies in a larger area are required for the maintenance of the epidemiological surveillance in the emphasized insular area.

Introdução: Leishmaniose Visceral (LV) é uma doença endêmica em municípios vizinhos à Ilha de Maré, situada na plataforma continental do município de Salvador, Bahia, Brasil. Entretanto, casos de LV não tem sido notificados nesta Ilha. Objetivo: O presente trabalho objetivou investigar a soroprevalência e sinais clínicos de Leishmaniose Visceral Canina (LVC) e identificar a população de flebótomos presentes no povoado de Botelho, Ilha de Maré. Metodologia: soro de cães locais foram testados para IgG anti-Leishmania por imunoensaios (triagem com teste rápido TR DPP™ e confirmatório com Elisa Indireto, Bio-Manguinhos/Fiocruz) e uma investigação entomológica foi conduzida para estimar e identificar a fauna flebotomínea da região. Resultados: sete de 106 amostras (6,6%) foram positivas usando o teste rápido. As amostras positivas foram encaminhadas ao Laboratório Central da Bahia para a confirmação por ELISA indireto. Entretanto, essas amostras apresentaram resultado negativo. Foram encontrados nove exemplares da espécie Pressatia choti, subfamília Phlebotominae, espécie frequentemente encontrada em áreas de transmissão de leishmaniose cutânea no Brasil. Conclusão: apesar deste trabalho não ter confirmado a presença de LVC na Ilha de Maré, novos inquéritos sorológicos e analises entomológicas em uma maior área são necessários para a manutenção de uma vigilância epidemiológica na região insular em destaque.

Attraction of the sandfly Lutzomyia longipalpis to possible biomarker compounds from dogs infected with Leishmania infantum.

Lutzomyia longipalpis (Diptera: Psychodidae) is the primary vector of Leishmania infantum (Kinetoplastida: Trypanosomatidae) in the Americas. Studies have been carried out to identify new alternatives for monitoring and controlling this sandfly species, particularly with the use of chemical baits. The attractiveness of odours emitted by foxes and alcohols found in some plants has already been demonstrated in laboratory tests with Lu. longipalpis. However, no studies have evaluated the responses of these insects to volatile organic compounds (VOCs) emitted by dogs. The present study was carried out to investigate the effects on Lu. longipalpis of individual and blends of VOCs identified in hair from dogs infected with L. infantum. Effects in male and female Lu. longipalpis were assessed using wind tunnel methodology. Individual compounds including octanal, nonanal, decanal and heptadecane showed capacity for activating and/or attracting male Lu. longipalpis. Only decanal and nonanal showed effects on females. The combination of octanal, decanal and heptadecane increased activation and attraction behaviour in male sandflies, as did the blend of octanal and decanal. These findings indicate that VOCs emitted by dogs may be an interesting source of new attractants of sandflies.

Occurrences of Phlebotomine Sand Flies (Diptera: Psychodidae) Potentially Associated with Leishmaniasis Transmission in Urban Parks in the City of São Paulo, Brazil.

A study of the phlebotomine sand flies was carried out between October 2011 and June 2013 in municipal parks in the city of São Paulo. A total of 173 specimens of 5 species were collected (Psychodopygus lloydi, Pintomyia fischeri, Lutzomyia amarali, Nyssomyia whitmani, and Migonemyia migonei). Three of these species may be involved in the transmission of the causative agent of tegumentary leishmaniasis. Surveillance and monitoring phlebotomine species from these areas are crucial as measure of prevention and control of leishmaniasis.

Detection of Leishmania infantum in Lutzomyia longipalpis captured in Campo Grande, MS / Detecção de Leishmania infantum em Lutzomyia longipalpis capturados em Campo Grande, MS

Leishmaniasis is a zoonotic disease caused by protozoa of the genus Leishmania (Ross, 1903) and is the focus of considerable attention in human and veterinary medicine. In the city of Campo Grande, MS, the causative agent of visceral leishmaniasis is Leishmania infantum (= L. chagasi) primary vector, comprising approximately 92.9% of the local sandfly population, is Lutzomyia longipalpis. The aim of this work was to compare real-time PCR with PCR as a tool for the detection of the kinetoplast DNA (kDNA) of L. infantum in sandflies. Sandflies of this species were caught, and a total of 38 samples with 1-4 individuals in each sample were obtained; these were distributed across 13 districts and divided between seven urban areas of the city of Campo Grande, MS. Three positive samples were found by PCR and, when using real-time PCR, this was able to detect the presence of this agent in 6 of the 13 districts sampled, which were all located on the outskirts of the city, where indicates the greater enzootic potential of these regions, as they are closer to natural forest reserves. We conclude that real-time PCR can be used for epidemiological studies of L. infantum.

A Leishmaniose é uma zoonose causada por protozoários do gênero Leishmania (Ross 1903), objetos de considerável atenção em medicina humana e veterinária. Na cidade de Campo Grande – MS, o agente etiológico da Leishmaniose Visceral é Leishmania infantum (= L. chagasi), e o principal vetor é a espécie Lutzomyia longipalpis, que representa cerca de 92,9% da população de flebotomíneos. Este trabalho teve como objetivo avaliar a PCR em tempo real como ferramenta para a detecção de kDNA de L. infantum em flebotomíneos, comparando-se com PCR convencional. Flebotomíneos dessa espécie foram capturados, somando 38 amostras de 1 a 4 espécimens cada, distribuídas em 13 bairros, divididos entre as 7 regiões urbanas da cidade de Campo Grande – MS, e armazenados a −70 °C até a extração de ADN e amplificação por PCR e PCR em tempo real. Das 38 amostras testadas, foram encontradas 3 amostras positivas pela PCR convencional e 11 pela PCR em Tempo Real. Na otimização da PCR em tempo real, a temperatura de dissociação do amplificado foi de 82, 89 °C. Neste estudo, utilizando-se a técnica da PCR em tempo real, foi possível detectar a presença desse agente em 6 dos 13 bairros amostrados, todos na periferia da cidade, indicando o maior potencial enzoótico dessas regiões, que têm maior proximidade com reservas de matas naturais. Conclui-se que a PCR em tempo real pode ser utilizada para estudo epidemiológico de L. infantum.

ASPECTS OF THE ECOLOGY OF PHLEBOTOMINES (Diptera: Psychodidae: Phlebotominae) IN AN AREA OF CUTANEOUS LEISHMANIASIS OCCURRENCE, MUNICIPALITY OF ANGRA DOS REIS, COAST OF RIO DE JANEIRO STATE, BRAZIL / Aspectos da ecologia dos flebotomineos (Diptera: Psychodidae: Phlebotominae) em area de ocorrencia de leishmaniose tegumentar, Municipio de Angra dos Reis, orla maritima do estado do Rio de Janeiro, Brasil

Over a complete two-year period, phlebotomine specimens were caught in an area of cutaneous leishmaniasis occurrence in the municipality of Angra dos Reis. A manual suction tube was used to catch phlebotomines on house walls, and also light traps in domestic and peridomestic settings and in the forest. This yielded 14,170 specimens of 13 species: two in the genus Brumptomyia and eleven in the genus Lutzomyia. L. intermedia predominantly in domestic and peridomestic settings, with little presence in the forest, with the same trend being found in relation to L. migonei, thus proving that these species have adapted to the human environment. L. fischeri appeared to be eclectic regarding location, but was seen to be proportionally more endophilic. L. intermedia and L. migonei were more numerous in peridomestic settings, throughout the year, while L. fischeri was more numerous in domestic settings except in March, April, May and September. From the prevalence of L. intermedia, its proven anthropophily and findings of this species naturally infected with Leishmania (Viannia) braziliensis, it can be incriminated as the main vector for this agent of cutaneous leishmaniasis in the study area, especially in the peridomestic environment. L. fischeri may be a coadjuvant in carrying the parasite.

Durante dois anos completos foram feitas capturas de flebotomíneos em área de leishmaniose tegumentar no município de Angra dos Reis. Utilizou-se tubo de sucção manual, para as capturas dos flebotomíneos pousados nas paredes da casa, além de armadilhas luminosas, no domicílio, peridomicílio e na mata. Foram obtidos 14.170 exemplares, de treze espécies, duas do gênero Brumptomyia França & Parrot 1921 e onze do gênero Lutzomyia França 1924. L. intermedia teve supremacia no peridomicílio e no domicílio, com pouca presença na mata, o mesmo ocorreu com L. migonei, comprovando a adaptação dessas espécies ao ambiente humano. L. fischeri aparece com característica eclética quanto ao local, mostrando-se proporcionalmente mais endófila. L. intermedia e L. migonei foram mais numerosas no peridomicílio, durante todos os meses do ano, enquanto L. fischeri, excetuando os meses de março, abril, maio e setembro, foi mais numerosa no domicílio. Pela prevalência, comprovada antropofilia e por ter sido encontrada infectada naturalmente por Leishmania (Viannia) braziliensis, L. intermedia pode ser incriminada como o principal vetor desse agente da leishmaniose tegumentar na área de estudo, sobretudo no ambiente peridomiciliar. L. fischeri, pelas características apresentadas, pode ser um coadjuvante na veiculação do parasita.

Aspectos da ecologia dos flebotomíneos (Diptera: Psychodidae: Phlebotominae) em área de ocorrência de leishmaniose tegumentar, município de Paraty, orla marítima do Estado do Rio de Janeiro, Brasil / Aspects of the ecology of phlebotomines (diptera: psychodidae: phlebotominae) in na area of cutaneous leishmaniasis occurrence, municipality of Paraty, coast of Rio de Janeiro State, Brazil

O crescente aumento dos casos de leishmaniose tegumentar, no município de Paraty, Estado do Rio de Janeiro, além da descrição do primeiro caso inédito e autóctone de leishmaniose cutâneo difusa, despertou o interesse de se realizar um estudo visando conhecer o perfil da fauna flebotomínica, estabelecendo a frequência domiciliar, mensal, além de analisar o índice de abundância das principais espécies. As capturas foram mensais, com permanência de quatro dias na área de estudo, durante três anos completos, outubro de 2009 a setembro de 2011 no bairro de São Gonçalo e outubro de 2011 a setembro de 2012 nos bairros de São Roque e Barra Grande. Os flebotomíneos foram capturados com tubos de sucção manual nas paredes internas e externas do domicílio, nos anexos de animais domésticos e na armadilha luminosa, modelo Shannon, armada no peridomicílio e na mata. As armadilhas luminosas CDC foram instaladas no intra, peridomicílio, margem e interior da mataForam obtidos 102.937 flebotomíneos, pertencentes a vinte e três espécies, três do gênero Brumptomyia França e Parrot, 1921 e vinte do gênero Lutzomyia França, 1924. Das espécies encontradas, seis já foram registradas com infecção natural: L. intermedia, L. fischeri, L. migonei, L. whitmani e L. pessoai por Leishmania braziliensis e L. ayrozai, por Leishmania naiffi. L. intermedia, em números absolutos, teve amplo predomínio sobre L. fischeri e L. migonei. Entretanto, o índice de abundância SISA mostrou L. fischeri como a primeira do ranking, sobretudo por ser abundante em todos os sítios de coleta. L. intermedia foi a segunda, pela abundância no ambiente domiciliar e com índices menores em relação à margem e a mata. L. migonei foi a terceira e L. whitmani a quarta mais abundante da fauna, particularmente na margem e no interior da mata...

The increasing cases of cutaneous leishmaniasis in the municipality of Paraty, Rio de Janeiro State, besides the description of the first new and autochthonous case of diffuse cutaneous leishmaniasis, aroused the interest of conducting a study to identify the characteristics ofphlebotomines establishing the frequency in domestic environments, monthly, besides analyzing the index of abundance of major species. Catches were monthly, with four daysremaining in the study area for three full years, October 2009 to September 2011 in São Gonçalo and October 2011 to September 2012 in the neighborhood districts of São Roque and Barra Grande. Phlebotomines were captured with manual suction tubes on internal andexternal walls of the house, annex of domestic animals and light trap, model Shannon, armed in peridomicile and inside the forest. The light traps, model CDC, were installed in domicile,peridomicile, borden and inside the forest. This yielded 102,937 phlebotomines belonging totwenty-three species, three genus Brumptomyia France e Parrot, 1921 and twenty of the genus Lutzomyia France, 1924. Six species were found with natural infection: L. intermedia,L. fischeri, L. migonei, L. whitmani e L. pessoai for Leishmania braziliensis and L. ayrozai for Leishmania naiffi. L. intermedia, in absolute numbers, had significant dominance of L.fischeri and L. migonei. However, the abundance index SISA showed L. fischeri as the first ranking, especially since it is abundant in all the study sites. L. intermedia was second for theabundance in the home environment and lower levels relative to the border and inside the forest. L. migonei was the third and L. whitmani the fourth most abundant fauna, particularlyin border and inside the forest...

Natural Leishmania sp. reservoirs and phlebotomine sandfly food source identification in Ibitipoca State Park, Minas Gerais, Brazil

Leishmania spp are distributed throughout the world and different species are associated with varying degrees of disease severity. However, leishmaniasis is thought to be confined to areas of the world where its insect vectors, sandflies, are present. Phlebotomine sandflies obtain blood meals from a variety of wild and domestic animals and sometimes from humans. These vectors transmit Leishmania spp, the aetiological agent of leishmaniasis. Identification of sandfly blood meals has generally been performed using serological methods, although a few studies have used molecular procedures in artificially fed insects. In this study, cytochrome b gene (cytB) polymerase chain reaction (PCR) was performed in DNA samples isolated from 38 engorged Psychodopygus lloydi and the expected 359 bp fragment was identified from all of the samples. The amplified product was digested using restriction enzymes and analysed for restriction fragment length polymorphisms (RFLPs). We identified food sources for 23 females; 34.8% yielded a primate-specific banding profile and 26.1% and 39.1% showed banding patterns specific to birds or mixed restriction profiles (rodent/marsupial, human/bird, rodent/marsupial/human), respectively. The food sources of 15 flies could not be identified. Two female P. lloydi were determined to be infected by Leishmania using internal transcribed spacer 1 and heat shock protein 70 kDa PCR-RFLP. The two female sandflies, both of which fed on rodents/marsupials, were further characterised as infected with Leishmania (Viannia) braziliensis. These results constitute an important step towards applying methodologies based on cytB amplification as a tool for identifying the food sources of female sandflies.

The occurrence of Phlebotomines (Diptera psychodidae) in a Leshmaniasis endemic area / Ocorrência de Flebotomíneos (Diptera psychodidae) em área endêmica de Leshmanioses

Objective: we describe the occurrence of phlebotomines in urban areas of Caxias, Maranhão state, whereautochthonous cases of visceral leishmaniasis (kala azar) and tegumentary leishmaniasis were reported.Method: The phlebotomines were captured in 40 districts in the periods January to December 2001, May toNovember 2004, and January to November 2005 using CDC light traps installed from 6 pm to 6 am in thehouses and peridomiciles. Results: a total of 28,336 specimens (21,006 males and 7,330 females) from 11species were collected. Ten of the species belonged to the genus Lutzomyia and one to the genus Brumptomyia.The most abundant species was Lutzomyia longipalpis (93.7%), which was also the most widely distributed inthe city, followed by L. whitmani (5.2%). Conclusions: this pattern of abundance indicates that these were thetwo species best adapted to the urban environment, and they were thus incriminated in the transmission of thecases of kala azar (Lutzomyia longipalpis) and tegumentary leishmaniasis (L. whitmani) reported in Caxias. Theother species were considered occasional or rare visitors to the areas

Objetivo: descreve-se a ocorrência de flebotomíneos em áreas urbanas com notificação de casosautóctones de leishmaniose visceral (calazar) e leishmaniose tegumentar. Método: nos períodos dejaneiro a dezembro de 2001, maio a novembro de 2004 e janeiro a novembro de 2005 foram feitosinquéritos entomológicos de flebotomíneos em 40 bairros da cidade de Caxias, estado do Maranhão.Os espécimes foram coletados utilizando-se armadilhas luminosas do tipo CDC instaladas das 18 às6 horas, no peri e intradomicílio. Resultados: foram coletados 28.336 espécimes (machos: 21.006;fêmeas: 7.330), de 11 espécies, sendo 10 pertencentes ao gênero Lutzomyia e 1 ao gêneroBrumptomyia. A espécie mais abundante foi L. longipalpis (93,7%) e também a mais bemdistribuída dentro da cidade, seguida por L. whitmani (5,2%). Conclusões: por esse padrão deabundância, verifica-se que essas duas espécies são as mais bem adaptadas ao ambiente urbano,sendo atraídas principalmente por galinheiros, chiqueiros e estábulos, além dos domicílioshumanos. As demais espécies foram consideradas visitantes ocasionais e raras nas áreas. Os casosde calazar e leishmaniose tegumentar notificados em Caxias são atribuídos a essas duas espécies,respectivamente.

Anthropogenic influence on the distribution, abundance and diversity of sandfly species (Diptera: Phlebotominae: Psychodidae), vectors of cutaneous leishmaniasis in Panama

In Panama, species of the genus Lutzomyia are vectors of American cutaneous leishmaniasis (ACL). There is no recent ecological information that may be used to develop tools for the control of this disease. Thus, the goal of this study was to determine the composition, distribution and diversity of Lutzomyia species that serve as vectors of ACL. Sandfly sampling was conducted in forests, fragmented forests and rural environments, in locations with records of ACL. Lutzomyia gomezi, Lutzomyia panamensis and Lutzomyia trapidoi were the most widely distributed and prevalent species. Analysis of each sampling point showed that the species abundance and diversity were greatest at points located in the fragmented forest landscape. However, when the samples were grouped according to the landscape characteristics of the locations, there was a greater diversity of species in the rural environment locations. The Kruskal Wallis analysis of species abundance found that Lu. gomezi and Lu. trapidoi were associated with fragmented environments, while Lu. panamensis, Lutzomyia olmeca bicolor and Lutzomyia ylephiletor were associated with forested environments. Therefore, we suggest that human activity influences the distribution, composition and diversity of the vector species responsible for leishmaniasis in Panama.

Phlebotomine fauna (Diptera: Psychodidae: Phlebotominae) in a residential area and in a fragment of savanna vegetation in the municipality of Pontal do Araguaia, Mato Grosso, Brazil / Fauna flebotomínica (Diptera: Psychodidae: Phlebotominae) em área residencial e em fragmento de cerrado no município de Pontal do Araguaia, Mato Grosso, Brasil

Identification of phlebotomine species in endemic areas is fundamental for analyzing the eco-epidemiological determinants of leishmaniasis. This study had the aim of investigating the phlebotomine fauna in an urban area and in a fragment of native savanna in the municipality of Pontal do Araguaia, State of Mato Grosso, Brazil, using CDC light traps. One hundred and twenty-three phlebotomine specimens belonging to seventeen different species were caught. Our results indicate synanthropic potential among vector species for leishmaniasis, such as the species Lutzomyia cruzi, L. sallesi and L. whitmani. The species L. cerradincola had never been recorded in this region, such that this is the first report of this species in the State of Mato Grosso.

A identificação das espécies de flebotomíneos em áreas endêmicas é fundamental para analisar os determinantes eco-epidemiológicos das leishmanioses. Este trabalho teve como objetivo verificar a fauna flebotomínica em área urbana e em fragmento de cerrado nativo no município de Pontal do Araguaia, Mato Grosso utilizando armadilhas luminosas tipo CDC. Foram capturados 123 flebotomíneos pertencentes a 17 espécies diferentes. Nossos resultados indicam potencial sinantrópico de espécies vetoras das leishmanioses, como as espécies Lutzomyia cruzi, L. sallesi e L. whitmani. A espécie L. cerradincola não havia sido registrada na região, sendo este o primeiro relato desta espécie no Estado de Mato Grosso.

Pesquisa de infecção natural de flebotomíneos por Leishmania, no Estado do Paraná / Research of natural infection of phlebotomines for Leishmania, in the State of Paraná

A leishmaniose tegumentar americana tem sido notificada em todos os estados do Brasil e no Paraná essa doença é endêmica. O objetivo deste trabalho foi detectar a infecção natural de flebotomíneos para verificar a competência vetorial destes insetos e a identificação da espécie parasitária. Os flebotomíneos foram coletados com armadilhas de Falcão e Shannon, nos municípios de Doutor Camargo, Fênix e Mandaguari, de novembro de 2005 a agosto de 2006. Coletaram-se 12.930 flebotomíneos, dos quais 2.487 fêmeas foram dissecadas e destes 1.230 fêmeas foram submetidas à reação em cadeia da polimerase. Pelo método da dissecação, foi detectada uma fêmea de Nyssomyia whitmani com infecção natural por flagelados e pela reação em cadeia da polimerase não se detectou a presença de DNA de Leishmania em nenhuma das fêmeas. Apesar de não ter sido detectada a infecção natural de Nyssomyia neivai nas localidades em apreço e ainda que os requisitos de incriminação vetorial não tenham sido atendidos, não se deve negligenciar o potencial vetorial desta espécie.

American cutaneous leishmaniasis has been reported in all Brazilian states and in the Paraná this disease is endemic. The objective of this work was to detect natural infections in phlebotomines to verify the vector competence of these insects and the identification of the parasite species. Phlebotomines were collected using Falcão and Shannon traps, in the municipalities of Doutor Camargo, Fênix and Mandaguari, between November 2005 and August 2006. from 12,930 phlebotomines were collected, 2,487 females were dissected and 1,230 dissected females had been submitted to polymerase chain reaction. Flagellates were detected in a female Nyssomyia whitmani that had been dissected and for polymerase chain reaction failed to detect Leishmania DNA in any females. Even though flagellates were not detected in Nyssomyia neivai it should still be considered as a potencial vector.